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. The largest and commercially appealing mineral depositscan be found in the abyssal sea floor of the Clarion-Clipperton Zone (CCZ), apolymetallic nodule province, in the NE Pacific Ocean, where experimentalmining is due to take place. In anticipation of deep-sea mining impacts, ithas become essential to rapidly and accurately assess biodiversity. For thisreason, ophiuroid material collected during eight scientific cruises fromfive exploration licence areas within CCZ, one area being protected from mining(APEI3, Area of Particular Environmental Interest) in the periphery of CCZand the DISturbance and re-COLonisation (DISCOL) Experimental Area (DEA),in the SE Pacific Ocean, was examined. Specimens were genetically analysedusing a fragment of the mitochondrial cytochrome c oxidase subunit I (COI).Maximum-likelihood and neighbour-joining trees were constructed, while fourtree-based and distance-based methods of species delineation (automatic barcode gap discovery, ABGD; barcode index numbers, BINs;general mixed Yule–coalescent, GMYC; multi-rate Poisson tree process, mPTP) were employed to propose secondary species hypotheses (SSHs)within the ophiuroids collected. The species delimitation analyses'concordant results revealed the presence of 43 deep-sea brittle star SSHs,revealing an unexpectedly high diversity and showing that the mostconspicuous invertebrates in abyssal plains have been so far considerablyunderestimated. The number of SSHs found in each area varied from five (IFREMER area) to 24 (BGR (Federal Institute for Geosciences and Natural Resources,Germany) area) while 13 SSHs were represented bysingletons. None of the SSHs were found to be present in all seven areas whilethe majority of species (44.2 %) had a single-area presence (19 SSHs).The most common species were Ophioleucidae sp. (Species 29), Amphioplus daleus (Species 2)and Ophiosphalma glabrum (Species 3), present in all areas except APEI3. The biodiversitypatterns could be mainly attributed to particulate organic carbon (POC) fluxes that could explain thehighest species numbers found in BGR (German contractor area) and UKSRL (UK Seabed Resources Ltd, UKcontractor area) areas. The five exploration contract areas belong to amesotrophic province, while conversely the APEI3 is located in anoligotrophic province, which could explain the lowest diversity as well asvery low similarity with the other six study areas. Based on these resultsthe representativeness and the appropriateness of APEI3 to meet its purposeof preserving the biodiversity of the CCZ fauna are questioned. Finally,this study provides the foundation for biogeographic and functional analysesthat will provide insight into the drivers of species diversity and its rolein ecosystem function.

Unexpected high abyssal ophiuroid diversity in polymetallic nodule fields of the northeast Pacific Ocean and implications for conservation